Regulatory Basics for Facility Design - PDF document

Regulatory Basics for Facility Design (WHO GMP): Biosafety Requirements Main Topics of the Presentation 2 • BSL in Laboratories (WHO Requirements) • Differences of BSL 1,2 and 3 in Laboratories and in Production F acilities • Decontamination by Fumigation • Air Supply Concept • Utilities Distribution and Effluent Treatment • Animal Facilities Risk Group for Laboratory Work 1 (1) • Risk Group 1: A microorganism that is unlikel

y to cause human or animal disease • Risk Group 2: A pathogen that can cause human or animal disease but is unlikely to be a serious hazard to laboratory workers, the community, livestock or environment • Risk Group 3: A pathogen that usually causes serious human or animal disease but does not ordinarily spread from one infected individual to another • Risk Group 4: A pathogen that usually causes serious human or animal disease and t

hat can be readily transmitted from on individual to another, directly or indirectly 3 1 WHO, Laboratory biosafety manual (3 rd edition) Risk Group for Laboratory Work 1 (2) 4 Risk Group Biosafety Level Laboratory Type Laboratory Practices Safety Equipment 1 Basic – BSL 1 Basic teaching, research Good microbiological techniques (GMT) Open work bench 2 Basic – BSL 2 Primary health and diagnostic services, research GMT + protective

clothing, biohazard sign Level 01 + BSC for potential aerosols 3 Containment – BSL 3 Special diagnostic services, research Level 2 + special clothing, controlled access, directional airflow BSC for all activities 4 Maximum containment – BSL 4 Dangerous pathogen units Level 3 + airlock entry, shower exit, special waste disposal Class III BSC, or positive pressure suits in conjunction with Class II BSCs, double - ended autoclave

, filtered air 1 WHO, Laboratory biosafety manual (3 rd edition) Requirements for Biosafety L evel (BSL) in Laboratories 5 BSL 1 BSL 2 BSL 3 BSL 4 Isolation a of laboratory No No Yes Yes Room sealable for decontamination No No Yes Yes Ventilation: - inward air flow - controlled ventilation system - HEPA - filtered air exhaust No Desirable Desirable No Yes Yes Yes/No b Yes Yes Yes Double - door entry No No Yes Yes Airlock No No No Yes Airlock w

ith shower No No No Yes Anteroom No No Yes - Anteroom with shower No No Yes/ No c No Effluent treatment No No Yes/ No c Yes Autoclave: - on site - in laboratory room - double - ended No Desirable No No Yes Desirable Desirable Yes Yes Yes Biological safety cabinets No Desirable Yes Yes Personnel safety monitoring capability d No No Desirable Yes a Environmental and functional isolation from general traffic. b Dependent on location of exhaust. c

Dependent on agent(s) used in the laboratory. d For example, window, closed - circuit television, two - way communication. Relevance of BSL for Production Facilities For the production of BSL relevant products, only the BSL 1 – 3 have to be taken into account. The risk of an BSL 4 production facility is too high. 6 Risk Assessment for Determination of BSL To provide the necessary level of safety for the specific work to be done, it has

to be evaluated based on a risk assessment 7 Assignment of BSL Consider… • t he used organism, (To which risk group belongs the used organism?) • the equipment, (Is the used equipment operated as an open system or is it a closed containment?) • a nd the procedure. (Are open procedures carried out in a biosafety cabinet or not?) 8 Differences of BSL 1, 2 and 3 in Laboratories and in Production Facilities 9 Features in Facilities WH

O CBC Concept Justification Isolation a of laboratory No Yes Automatically implemented because of the WHO GMP guidelines ( Isolation of production rooms through separate airlocks , leading into separate corridors) Room sealable for decontamination No Yes (if required) To avoid cross - contaminations in a facility in which more than one product or strain is produced Ventilation: - inward air flow (differential pressure) - controlled ventilation

system (air recirculation) - HEPA - filtered air exhaust No No (Possible ) No Yes/No (inward air flow) Yes No - Depending on the risk of the product c - Air recirculation allow if HEPA - filtered and recirculated within the same bio - positive production rooms Double - door entry No Yes Automatically implemented by the separation of the different clean rooms grades and the design of ALs between them Airlock No Yes Automatically implemented

by the separation of the different clean rooms grades and the design of ALs between them Airlock with shower No No Anteroom No Yes Automatically implemented (d esigned as airlocks, see above) Anteroom with shower No No Effluent treatment No Yes Because of the size of production volume and therefore also waste water volume (can be more than several 100 litres) Autoclave: - on site - in laboratory room - double - ended No Yes Yes Yes Double -

ended autoclaves facilitate the process of handling contaminated goods / material. No wrapping to bring goods out of the bio+ area to the cleaning area. Cleaning area directly connected Biological safety cabinets No Yes/No Depending on the risk of the produced product d Personnel safety monitoring capability b No Yes /No Decide by the customer a Environmental and functional isolation from general traffic. b For example, window, closed - ci

rcuit television, two - way communication. c Differential pressure can either be positive, 0 or negative, depending on the risk, (e.g. if the risk of reverse mutation of t he attenuated strain to the wild type strain exists, the building should have negative differential pressure to avoid the release of the virus into the environment) d If the risk of reverse mutation of the attenuated strain exists, BSCs should be installed Difference BSL 1:

WHO and CBC Design Concept 10 11 Difference BSL 2: WHO and CBC Design Concept Features in Facilities WHO CBC Concept Justification Isolation a of laboratory No Yes Automatically implemented because of the WHO GMP guidelines ( Isolation of production rooms through separate airlocks , leading into separate corridors) Room sealable for decontamination No Yes To avoid cross - contaminations in a facility in which more than one product or strain i

s produced. Decontamination before maintenance if pathogens are handled Ventilation: - inward air flow (differential pressure) - controlled ventilation system (air recirculation) - HEPA - filtered air exhaust Desirable Desirable ( Possible 1 ) No Yes/No (inward air flow) Yes Yes - Depending on the risk of the product c - Air recirculation allowed if HEPA - filtered and recirculated within the same bio - positive production rooms - Directly in

tegrated on the clean room wall or centralized in the air handling unit system d Double - door entry No Yes Automatically implemented by the separation of the different clean rooms grades and the design of ALs between them Airlock No Yes Automatically implemented by the separation of the different clean rooms grades and the design of ALs between them Airlock with shower No No Possible if emergency showers are needed Anteroom No Yes Automatic

ally implemented (d esigned as airlocks, see above) Anteroom with shower No No Effluent treatment No Yes Because of the size of production volume and therefore also waste water volume (can be more than several 100 litres) Autoclave: - on site - in laboratory room - double - ended Desirable No No Yes Yes Yes Double - ended autoclaves facilitate the process of handling contaminated goods / material. No wrapping to bring goods out of the bio+ ar

ea to the cleaning area. Cleaning area directly connected Biological safety cabinets Desirable Yes Because of the large amount of the produced product e Personnel safety monitoring capability b No Yes /No Decide by the customer a Environmental and functional isolation from general traffic. b For example, window, closed - circuit television, two - way communication. c Differential pressure is either 0 or negative, for mAb facilities the di

fferential pressure is positive d Consequence of centralized HEPA filter  pipe system is also contaminated e if process can’t be performed in an BSC, customer - made UAF with inward air flow are used. For mAb and genetically modified products not necessary Features in Facilities WHO CBC Concept Justification Isolation a of laboratory Yes Yes Room sealable for decontamination Yes Yes Ventilation: - inward air flow (differential pressure) -

controlled ventilation system (air recirculation) - HEPA - filtered air exhaust Yes Yes (Possible) Yes/No b Yes (inward air flow) Yes Yes - N egative pressure in bio - positive areas to avoid the flow of contaminated air into the clean bio - negative area of production - Air recirculation allow if HEPA - filtered and recirculated within the same bio - positive production rooms - Directly integrated on the clean room wall Double - door entry

Yes Yes Airlock No Yes Automatically implemented by the separation of the different clean rooms grades and the design of ALs between them Airlock with shower No Yes Because of the large amount of produced product a utomatically implemented by the separation of the different clean rooms grades and the design of ALs between them Anteroom Yes Yes Anteroom with shower Yes/ No c Yes Automatically implemented (d esigned as airlocks with shower, se

e above) Effluent treatment Yes/ No c Yes On site liquid decontamination Autoclave: - on site - in laboratory room - double - ended Yes Desirable Desirable Yes Yes Yes Double - ended autoclaves facilitate the process of handling contaminated goods / material. No wrapping to bring goods out of the bio+ area to the cleaning area. Cleaning area directly connected Biological safety cabinets Yes Yes Personnel safety monitoring capability d Desira

ble Yes 12 Difference BSL 3: WHO and CBC Design Concept a Environmental and functional isolation from general traffic. b Dependent on location of exhaust. c Dependent on agent(s) used in the laboratory. d For example, window, closed - circuit television, two - way communication. Conclusion for Facilities (BSL 1 - 3) 13 • Generally over - designed compared to laboratories because of the large amount of product which has to be handled • B

SL 1 and 2: the range of risks is huge (depends on the product)  e.g. i n Europe, genetically modified microorganisms are BSL 2 because of the risk of release to the environment, not because of pathogentic aspects  On the other hand, if e.g. the risk of reverse mutation exists for attenuated virus strains, a BSL 1 production facility may need additional biosafety features normally not required for BSL 1 (e.g. the use of biosafety cab

inets, etc.) • At the end, critical BSL 1 and 2 production facilities may not be that different a nymore compared to BSL 3 laboratories / facilities. 14 Bio - positive Production Rooms Bio - negative Production Rooms Airlocks VHP Airlock BSL 1/2: Production  pressure in the bio - positive area ≤0 (potentially pathogenic product) Grade C Grade D Grade B Grade CNC Air flow +15 Supply Corridor 15 Barrier Airlocks BSL 1/2: Air flow

and barrier a irlocks for protection of the different states of the product Grade C Grade D Grade B Grade CNC Air flow +15 Supply Corridor 16 BSL 1/2: D ouble - ended autoclaves (into and out of the bio - positive area) Grade C Grade D Grade B Grade CNC Decontamination of used Items 17 WHO Requirement for BSL 1 and 2: “All contaminated materials, specimens and cultures must be decontaminated before disposal or cleaning for reus

e .” Bio - negative area Bio - positive area (BSL 2) WHO, Laboratory biosafety manual (3 rd edition), chapter 3, Laboratory working areas, Point 3 Grade D Grade CNC Decontamination of used Items 18 Use of VHP MALs (fumigation c hamber) for… 1. …the surface decontamination of material from the bio - positive to the bio - negative area. 2. …for surface sterilization during the transport of material / product between different clean r

oom grades. Further explanations will follow in few slides Bio - negative area Bio - positive area (BSL 2) Grade D Grade CNC 19 Bio - positive Production Rooms Bio - negative Production Rooms Bio - negative Production Rooms Bio - positive in case of disaster Grade C Grade D Grade B Grade CNC BSL 3 Bio - Positive in case of Disaster 20 “Bio - positive in case of disaster” rooms are intended for quarantine purpose. • Designed as bio - posi

tive rooms but operated as bio - negative rooms • In case of disaster with quarantined virus / bacteria solutions (during / after inactivation), the rooms become bio - positive and will be decontaminated appropriately afterwards  Decontamination shall only be done with a mobile fumigation system (  see VHP decontamination in few slides) 21 Grade C Grade D Grade B Grade CNC BSL 3: ALs Airlocks 22 BSL 3: Air Flow in ALs Barrier A

irlocks which are mandatory Grade C Grade D Grade B Grade CNC Air flow Barrier Airlocks by chance Airlocks with Shower in BSL 3 Facilities 23 WHO Requirement: Anteroom with shower for BSL 3 laboratories required or not required (depending on agents). 1 CBC Design Concept : • Airlock with shower available on the way out of the bio - positive production rooms • Waste water is decontaminated (explained later in few slides) • Shower

designed as grade D because for this grade no monitoring is required for “in operation” WHO, Laboratory biosafety manual (3 rd edition), chapter 1 Grade C Grade D Grade CNC Air flow WHO Requirement: Special attention should be taken to processes causing safety problems, like formation of aerosols (centrifugation, opening of containers,…)  Use of Biological Safety Cabinets (BSC ) (also for BSL 1 and 2 if necessary) e.g. d

uring the chromatography  if the different fractions of the product are collected in open bottles , this has to be done in a BSC (including the pooling). The chromatography column can be outside of the BSC (if sterile bags are taken instead, they can be connected aseptically and no BSC would be needed �= closed system ) BSCs in Biosafety Relevant Production Facilities 24 WHO, Laboratory biosafety manual (3 rd edition), chapt

er 3 , Essential biosafety equipment, Point 2 Bio - positive in case of disaster Bio - positive area Decontamination by Fumigation 25 Decontamination by Fumigation 26 1 WHO, Laboratory biosafety manual (3 rd edition), chapter 4 , Laboratory design and facilities, Point 7 WHO requirement: “ The laboratory room must be sealable for decontamination. Air - ducting systems must be constructed to permit gaseous decontamination.” 1  Remember

! Air - tightness of clean rooms was the topic of the previous presentation  Usage of vaporized H 2 O 2 + less toxic than formalin and EtO (which are carcinogen) + not flammable or explosive in the used concentration + short fumigation cycles + decomposition into water and oxygen + no residues on clean room surfaces - higher investment costs - stronger precautions to avoid damages to the facility and the personnel Mobile VHP Decontamina

tion System 27 Example of an (schematic) mobile VHP decontamination system for an VHP airlock (e.g. used for the transport between bio - positive and bio - negative area) Mobile VHP generator VHP airlock Centralized VHP Decontamination System for Entire Rooms or Facilities 28 • Off - the - shelf, but still has to be integrated into the HVAC system which requires customized and high qualification effort • Programming of fully automat

ed cycles • One fumigation cycle for a large facility • Additional computerized control system is needed • Higher price than mobile systems • Higher safety risk than mobile systems 29 Example for a complex centralized VHP decontamination system 30 Example for a complex centralized VHP decontamination system 31 Example for a complex centralized VHP decontamination system 32 Example for a complex centralized VHP decontamination sys

tem 33 Example for a complex centralized VHP decontamination system Air Supply Concept 34 Air F low S ystem 35 WHO Requirement for BSL 1 and 2: “In the planning of new facilities, consideration should be given to the provision of mechanical ventilation systems that provide an inward flow of air without recirculation.” 1 BUT for BSL 3: “Air may be high - efficiency particulate air (HEPA) filtered, reconditioned and recircul

ated within that laboratory .” 2 CBC Design Concept: Recirculating air flow with HEPA filter for outflow (only in BSL 2 and BSL 3 rooms) and inflow (BSL 1, 2 and 3). 1 WHO, Laboratory biosafety manual (3 rd edition), chapter 3, Design features, Point 15 2 WHO, Laboratory biosafety manual (3 rd edition), chapter 4 , Laboratory design and facilities, Point 8 So… Implementation in the Design of the Bio+ Clean Rooms 36 Incoming HEPA - filt

ered recirculated air (BSL 1, 2 and 3) HEPA - filtered e xhaust and air for recirculation (BSL 2 and 3) 1 WHO, Laboratory biosafety manual (3 rd edition), chapter 3, Design features, Point 15 2 WHO, Laboratory biosafety manual (3 rd edition), chapter 4 , Laboratory design and facilities, Point 8 Utilities Distribution and Effluent Treatment 37 General Requirements (1) 38 WHO R equirement: 1. “A dependable supply of good quality water is

essential. There should be no cross - connections between sources of laboratory and drinking - water supplies . An anti - backflow device should be fitted to protect the public water system .” WHO , Laboratory biosafety manual (3rd edition), chapter 3, Procedures, Point 16 2. “Water from the personnel shower and toilet may be discharged directly to the sanitary sewer without treatment .” WHO , Laboratory biosafety manual (3rd e

dition), chapter 5, Laboratory design and facilities, Point 4 3. “Effluents should be collected in closed vessels for further autoclaving and/or disposal .” WHO , Laboratory biosafety manual (3 rd edition), chapter 12, Automated equipment ( sonicators , vortex mixers), Point 2 General Requirements (2) 39 WHO Requirement : 4. “ Water - treatment plants and distribution systems should be designed, constructed and maintained so as t

o ensure a reliable source of water of an appropriate quality . ” WHO TRS 961, Annex 6, paragraph 12.6 5. “ An effluent treatment system may be required , depending on the risk assessment for the agent(s) being handled .” WHO, Laboratory biosafety manual (3 rd edition), chapter 3, Design features, Point 7 6. “ Backflow - precaution devices must be fitted to the water supply .” WHO, Laboratory biosafety manual (3 rd edition), ch

apter 4, Laboratory design and facilities, Point 13 Conclusion for the CBC Design Concept 40 As a result of the above mentioned points (and other requirements 1 ), the CBC Design Concept implements the following points for clean room facilities: • PW is generated out of pretreated/softened water by reverse osmosis and electro - deionization (EDI) �= satisfies requirement 4 on the slide before • WFI is generated out of PW by disti

llation �= satisfies requirement 4 • Separate loops for bio - positive and bio - negative areas (for PW and for WFI) �= 1/6 • Bio - positive loops are supplied by bio - negative loos (for PW and for WFI) �= 1/6 • Each bio - positive area has its own bio - positive PW and WFI loop� = 1/6 • An automatic decontamination system is propo�sed = 3/5 • Annotation: loop distribution system do normally not use

backflow - prevention valves (would make no sense). Separation is achieved by separated loops / subloops as explained above 1 WHO, TRS 970, Annex 2: Water for Pharmaceutical Use 41 Bio - negative Loops Bio - positive Loops Water purification CBC design concept for U tility D istribution (PW, WFI and PS) 42 CBC design concept for W aste Water Treatment • Three different products (bio+ areas) • No difference between BSL 1, 2 or 3 wa

ste water • Each bio+ area has its own waste water collection tank (permitted independence and precaution of cross - contamination) • System allows continuous decontamination Animal Facilities 43 Animal Facility Containment Levels 44 Risk Group Containment Level Laboratory Practices and Safety Equipment 1 ABSL - 1 Limited access, protective clothing and gloves 2 ABSL - 2 ABSL - 1 practices + hazard warning signs. Class I or Class II

BSCs for activities that produce aerosols. Decontamination of waste and cage before washing 3 ABSL - 3 ABSL - 2 practices + controlled access. BSCs and special protective clothing for all activities 4 ABSL - 4 ABSL - 3 practices + strictly limited access. Clothing change before entering. Class III BSCs or positive pressure suits. Shower on exit. Decontamination of all wastes before removal from facility ABSL = animal facility biosafety lev

el WHO, Laboratory biosafety manual (3 rd edition), chapter 6 ABSL - 1 45 WHO Requirement : • For maintenance of most stock animals after quarantine • For animals that are inoculated with agents (Risk group 1 ) • Supportive operations related to the animal holding (cage washing, transport of material and equipment which were in contact with the animals, and transport of animals) CBC Design Concept: • Supply and return corridor (both A

BSL - 1) • ABSL - 1 animal rooms between the supply / return corridors • Unidirectional path of material/personnel/animals • Airlocks with lockers to put on dedicated garments for ABSL - 1 area WHO, Laboratory biosafety manual (3 rd edition), chapter 6 , Animal facility – Biosafety Level 1 ABSL - 1 46 Supply corridor Return corridor ABSL 1 Animal/Personnel/Material Flow ABSL - 2 47 WHO Requirement: • For animals that are inoculated

with agents (Risk group 2) • Decontamination of waste material, bedding, animal cage by an autoclave • BSCs for work that may generate aerosols • Quarantine area for the animals CBC Design Concept : • Supply (ABSL - 1), bio - negative (ABSL - 1), bio - positive (ABSL - 2) and return corridor (ABSL - 2) • ABSL - 2 room between the bio - positive and bio - negative corridor • Unidirectional path of material/personnel/animals • Ai

rlocks entering ABSL - 2 rooms out of bio - negative corridor • Quarantine is designed as ABSL - 2 rooms • Access through airlocks only with changing of garment • Decontamination of all waste and materials which leaves the ABSL - 2 area • Use of BSC for work which may generate aerosols (inoculation, bleeding, etc.) WHO, Laboratory biosafety manual (3 rd edition), chapter 6 , Animal facility – Biosafety Level 2 48 Supply corridor Ret

urn corridor Bio+ corridor Bio - corridor ABSL 1 Animal/Personnel/Material Flow ABSL 2 Decontamination via autoclave and steam chamber CBC design concept example for Q uarantine Area 49 Supply corridor ABSL 1 ABSL 2 Animal reception Animal Facilities: Design of S upportive A reas 50 51 ABSL 1 Grade D Washing Area • After decontamination via autoclave (only out of ABSL 2 area), material is brought to the washing area (via return corri

dor) • Pass - through washing machines wash the dirty material • Clean material can be distributed in the facility again by the supply corridor Grade D area because in this animal house other procedure were done in which clean room clothes were needed Material Flow Similarities and Special Case 52 Similarities: • As for BSL 2 area, waste water from ABSL - 2 is collected and decontaminated • As for BSL 1 and 2, ABSL - 2 area ha

s its own water supply loop  ABSL - 1 doesn’t need a separate loop within the animal house, because inactive agents are handled. The ABSL - 1 is needed because of the general handling of animals Special Case: • No recirculation of air (because of the smell of the animals)  Centralized air handling units for conditioning of outside air, air supply to all areas of the building  Centralized exhaust air handling unit  Segregation o

f ABSL - 2 area from ABSL - 1 and black (unclassified) area by terminal HEPA filters for supply and exhaust air in the A BSL 2 rooms  Negative differential pressure in the animal holding rooms, ABSL - 1 (smelly air) and ABSL - 2 (biosafety containment) 53 CBC design concept example for a V entilation System Centralized supply and exhaust air handling unit 54 Detailed Room Typicals for ABSL - 1 and ABSL - 2 Terminal HEPA filter in ABSL -